A label free exonuclease III-aided fluorescence assay for adenosine triphosphate based on graphene oxide and ligation reaction

Zhu, Wenqing; Zhao, Ziwei; Li, Zhen; Li, Hong; Jiang, Jian‐Hui; Shen, Guo‐Li; Yu, Ru‐Qin

doi:10.1039/c2nj41055a

Cited by 22 publications

(13 citation statements)

References 39 publications

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“…It indicated that Exo III cleavage enhanced 270% S/B ratio in comparison with that of Hinf I‐digested DNA–GO. Furthermore, it is known that Exo III had been used to enhance fluorescence intensity strongly for enzyme assays except restriction endonuclease assay . It demonstrates that Exo III‐aided fluorescence assay can be used for the detection of restriction endonuclease activity in the presence of GO, leading to high S/B ratio.…”

Section: Resultsmentioning

confidence: 99%

Simple and Sensitive Fluorescence Assay of Restriction Endonuclease on Graphene Oxide

Gang

2015

Bulletin Korean Chem Soc

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Restriction endonucleases hydrolyze internal phosphodiester bonds at specific sites in a DNA sequence. These enzymes are essential in a variety of fields, such as biotechnology and clinical diagnostics. It is of great importance and necessity for the scientific and biomedical use of enzymes to measure endonuclease activity. In this study, graphene oxide (GO) has been used as a platform to measure enzyme activity with high sensitivity. To increase the detection sensitivity of Hinf I, the endonuclease‐digested reaction was treated with exonuclease III (Exo III) and a fluorescence assay was conducted to measure the emission. Results showed that Exo III treatment enhanced 2.7‐fold signal‐to‐background ratio for the detection of Hinf I compared with that done without Exo III in the presence of GO.

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Section: Resultsmentioning

confidence: 99%

Simple and Sensitive Fluorescence Assay of Restriction Endonuclease on Graphene Oxide

Gang

2015

Bulletin Korean Chem Soc

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“…13, 14 The method is based on the finding by Ono and Togashi that Hg 2+ can selectively bind between two mutually facing thymine (T) bases to form T-Hg 2+ -T complexes. [21][22][23][24] Recently, our group has successfully detected specific ssDNA oligomer sequences and proteins with high sensitivity and selectivity using AFM-based single-molecule force spectroscopy (SMFS). 16 19 Recently, Zhang and co-workers reported a surface enhanced resonance Raman scattering (SERRS) method for Hg 2+ detection by using nanoporous gold as a substrate and a Cy5-labeled aptamer as an optical tag.…”

Section: Introductionmentioning

confidence: 99%

“…Indeed, many efforts have been made to develop label-free methods for detecting small molecules. [21][22][23][24] Recently, our group has successfully detected specific ssDNA oligomer sequences and proteins with high sensitivity and selectivity using AFM-based single-molecule force spectroscopy (SMFS). 24 We have proven that SMFS can be effectively used as a label-free sensing method for oligonucleotides with a detection limit of 1 nM, in particular distinguishing between complementary and mis-matched ssDNA sequences.…”

Section: Introductionmentioning

confidence: 99%

A novel aptasensor based on single-molecule force spectroscopy for highly sensitive detection of mercury ions

Michaelis

Wei

et al. 2015

Analyst

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We have developed a novel aptasensor based on single-molecule force spectroscopy (SMFS) capable of detecting mercury ions (Hg(2+)) with sub-nM sensitivity. The single-strand (ss) DNA aptamer used in this work is rich in thymine (T) and readily forms T-Hg(2+)-T complexes in the presence of Hg(2+). The aptamer was conjugated to an atomic force microscope (AFM) probe, and the adhesion force between the probe and a flat graphite surface was measured by single-molecule force spectroscopy (SMFS). The presence of Hg(2+) ions above a concentration threshold corresponding to the affinity constant of the ions for the aptamer (about 5 × 10(9) M(-1)) could be easily detected by a change of the measured adhesion force. With our chosen aptamer, we could reach an Hg(2+) detection limit of 100 pM, which is well below the maximum allowable level of Hg(2+) in drinking water. In addition, this aptasensor presents a very high selectivity for Hg(2+) over other metal cations, such as K(+), Ca(2+), Zn(2+), Fe(2+), and Cd(2+). Furthermore, the effects of the ionic strength and loading rate on the Hg(2+) detection were evaluated. Its simplicity, reproducibility, high selectivity and sensitivity make our SMFS-based aptasensor advantageous with respect to other current Hg(2+) sensing methods. It is expected that our strategy can be exploited for monitoring the pollution of water environments and the safety of potentially contaminated food.

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“…Recently, biosensors based on nanomaterials have been reported for the detection of protease, nucleic acids, metal ions and small molecules. [25][26][27][28] In this paper, we construct a DNAzyme-AuNPs based biosensing system for the highly sensitive and selective detection of Pb 2+ ions, by taking advantage of the super fluorescence quenching efficiency of the AuNPs. 29 A classic Pb 2+ -dependent GR-5 DNAzyme was chosen as the catalytic unit instead of 8-17 DNAzyme, because GR-5 DNAzyme has been reported to offer a much higher selectivity towards Pb 2+ ions than 8-17 DNAzyme over other competing metal ions.…”

Section: Introductionmentioning

confidence: 99%

“…29 A classic Pb 2+ -dependent GR-5 DNAzyme was chosen as the catalytic unit instead of 8-17 DNAzyme, because GR-5 DNAzyme has been reported to offer a much higher selectivity towards Pb 2+ ions than 8-17 DNAzyme over other competing metal ions. 24,28,30 In the absence of Pb 2+ ions, the FAM-labeled GR-5 DNAzyme was in close proximity to the surface of AuNPs and the fluorescence signal was quenched. In the presence of Pb 2+ ions, the GR-5 DNAzyme was activated and cleaved the substrate strand at the RNA site (rA) into two parts, which led to a short FAM-linked oligonucleotide fragment releasing from the surface of the AuNPs, and the fluorescence intensity gradually recovered.…”

Section: Introductionmentioning

confidence: 99%

A highly sensitive and selective biosensing strategy for the detection of Pb2+ ions based on GR-5 DNAzyme functionalized AuNPs

Wang

Huang

et al. 2013

New J. Chem.

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A label free exonuclease III-aided fluorescence assay for adenosine triphosphate based on graphene oxide and ligation reaction

Cited by 22 publications

References 39 publications

Simple and Sensitive Fluorescence Assay of Restriction Endonuclease on Graphene Oxide

Simple and Sensitive Fluorescence Assay of Restriction Endonuclease on Graphene Oxide

A novel aptasensor based on single-molecule force spectroscopy for highly sensitive detection of mercury ions

A highly sensitive and selective biosensing strategy for the detection of Pb2+ ions based on GR-5 DNAzyme functionalized AuNPs

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